Simultaneous determination of praziquantel, pyrantel embonate, febantel and its active metabolites, oxfendazole and fenbendazole, in dog plasma by liquid chromatography/mass spectrometry.

A liquid chromatography-electrospray-mass spectrometry method (LC/MS) has been developed and validated for determination of praziquantel (PZQ), pyrantel (PYR), febantel (FBT), and the active metabolites fenbendazole (FEN) and oxfendazole (OXF), in dog plasma, using mebendazole as internal standard (IS). The method consists of solid-phase extractions on Strata-X polymeric cartridges. Chromatographic separation was carried out on a Phenomenex Gemini C6 -Phenyl column using binary gradient elution containing methanol and 50 mm ammonium-formate (pH 3). The method was linear (r(2) ≥ 0.990) over concentration ranges of 3-250 ng/mL for PYR andFEB, 5-250 ng/mL for OXF and FEN, and 24-1000 ng/mL for PZQ. The mean precisions were 1.3-10.6% (within-run) and 2.5-9.1% (between-run), and mean accuracies were 90.7-109.4% (within-run) and 91.6-108.2% (between-run). The relative standard deviations (RSD) were <9.1%. The mean recoveries of five targeted compounds from dog plasma ranged from 77 to 94%.The new LC/MS method described herein was fully validated and successfully applied to the bioequivalence studies of different anthelmintic formulations such as tablets containing PZQ, PYR embonate and FBT in dogs after oral administration.

[Analysis of laboratory data of 155 patients with pheochromocytoma-paraganglioma syndrome diagnosed during the past 20 years]. / A laboratóriumi diagnosztika eredményei az elmúlt 20 évben kórismézett 155 phaeochromocytoma/paraganglioma szindrómás beteg adatainak elemzése alapján.

INTRODUCTION: Laboratory diagnosis of pheochromocytoma-paraganglioma syndrome has been markedly improved during the past two decades. AIM: Retrospective assessment of diagnostic utility of urinary catecholamines and their metabolites as well as serum chromogranin A in 155 patients diagnosed at the 2nd Department of Medicine, Semmelweis University. METHOD: Urinary catecholamines and metabolites were measured using high-performance liquid chromatography with electrochemical detection in 155 patients with pheochromocytoma-paraganglioma (of whom 28.4% had hereditary background) and in 170 non-pheochromocytoma patients used as controls. Serum chromogranin A was measured by immunoradiometry. RESULTS: Sensitivity (93.2%) and specificity (87.0%) of urinary fractionated metanephrines were higher than those of urinary catecholamines (90.9% vs. 85.7%, respectively) and serum chromogranin A (88.7% and 77.5%, respectively). Urinary normetanephrine and serum chromogranin A correlated positively with tumor size (r = 0.552, p<0.0001 and r = 0.618, p<0.0001, respectively). CONCLUSIONS: These data confirm the diagnostic utility of urinary catecholamines and their metabolites. Urinary normetanephrine and serum chromogranin A may help to estimate tumour mass and probably tumour progression.

[Hair analysis of abused drugs with gas-chromatography mass spectrometry]. / Abúzusszerek hajból történo meghatározása GC/MS módszerrel.

Beside the traditionally used body-fluids, defining the abuse-material by the use of hair samples is more and more widespread in the forensic toxicological practice. Using the hair allows the rectrospective examination of the abuse-material, and due to the sensitive measuring technics, even one-time use can be proven. A further possibility is the segment-analysis which allows investigation of the abuse-history retroactive for months depending on the length of the hair. The quantitative parameters of the abuse can not always be estimated precisely since the details of the build-up in the hair are complicated and are not clear even today. Furthermore, the sampling, sample preparation and the measuring method will all influence the results. Our paper reviews the opiates, cocain, amfetamin derivatives, cannabinoids, alcohol-consumption markers and the frequently found drugs in the forensic toxicology as determined by using hair samples.

[Hair analysis of abused and therapeutic drugs in forensic toxicology]. / Gyógyszerek és abúzusszerek igazságügyi toxikológiai vizsgálata hajból.

Hair analysis for abused drugs has been gaining increasing significance in forensic sciences. Hair is a special matrix for the retrospective investigation of chronic drug abuse or poisoning in criminal cases and allows to demonstrate with sensitive methods even a single administration in low amount. Segmental hair analysis can yield the information about the time course of the substance use. The background of drug incorporation mechanism is not yet understood in full details and cannot be evaluated exactly in all cases. The hair sampling, sample preparation, analytical performance are very important for final results. The outcomes of hair analysis have been reviewed by dividing into six groups: opiates, cocaine, amphetamines, cannabinoids, abused therapeutic drugs and the markers of chronic alcohol consumption.

[Determination of ethanol concentrations from bile and its importance in forensic medicine]. / Epébol történo etilalkohol meghatározás jelentosége az igazságügyi orvostani gyakorlatban.

Determination of post-mortem ethanol concentration is a very important analytical examination in forensic toxicological daily routine. But in many cases, blood and urine are not available in appropriate quality and quantity for the analysis. In these cases, other biological matrices such as liquor vitreous humor, saliva, bile, etc. can be used for ethanol concentration determination. In toxicological laboratories, these kinds of examinations have been done for decades. In our institute, we compared the ethanol concentrations of blood, urine and bile samples collected from 115 autopsies. Determination of ethanol levels was carried out by using headspace GC according to international practice. The average of ethanol concentration ratios ([blood]/[bile]) was 1,220. Furthermore, we examined the pH of bile samples, but we did not find any relationships between ethanol levels and pH values. In 90 percent of results, positive correlation can be shown between bile and blood ethanol levels. In remaining cases, we tried to account for the reasons of differences by looking into autopsy reports.

Frequency and structure of stimulant designer drug consumption among suspected drug users in Budapest and South-East Hungary in 2012-2013.

Identification of abuse and frequency patterns of stimulant designer drugs (SDDs) provides important information for their risk assessment and legislative control. In the present study urine and/or blood samples of suspected drug users in criminal cases were analysed by GC-MS for 38 SDDs, and for the most frequent illicit and psychoactive licit drugs in Hungary. Between July 2012 and June 2013, 2744 suspected drug users were sampled in Budapest and during 2012 and 2013, 774 persons were sampled in South-East Hungary (Csongrád County - neighbour the Romanian and Serbian borders). In Budapest 71.4% of cases, and in South-East Hungary 61% of cases were positive for at least one substance. Pentedrone was the most frequent SDD in both regions; however, the frequency distribution of the remaining drugs was highly diverse. SDDs were frequently present in combination with other drugs - generally with amphetamine or other stimulants, cannabis and/or benzodiazepines. The quarterly distribution of positive samples indicated remarkable seasonal changes in the frequency and pattern of consumption. Substances placed on the list of illicit drugs (mephedrone, 4-fluoro-amphetamine, MDPV, methylone, 4-MEC) showed a subsequent drop in frequency and were replaced by other SDDs (pentedrone, 3-MMC, methiopropamine, etc.). Newly identified compounds from seized materials were added to the list of new psychoactive substances ("Schedule C"). While the risk assessment of substances listed in Schedule C has to be performed within 2 years after scheduling, continuous monitoring of their presence and frequency among drug users is essential. In summary, our results suggest which substances should be dropped from the list of SDDs measured in biological samples; while the appearance of new substances from seized materials indicate the need for developing adequate standard analytical methods.

Evaluation of a fatal propofol intoxication due to self administration.

Propofol is a short acting intravenous anaesthetic, active agent of Diprivan. The abuse potential of propofol has not completely defined, but there are anecdotal case reports in the literature about propofol abuse and dependency. This report presents a fatal case of a middle age female victim who died of self-administered propofol intoxication. The propofol level of the blood sample was measured with gas chromatography-mass spectrometry after liquid-liquid extraction. The results of toxicological investigation suggested that death was not directly caused by propofol intoxication, however, based on the pathomorphological changes detected during the medico-legal autopsy we supposed that the fatal outcome was resulted by respiratory depression after rapid injection.

Alcohol-related mortality risk in natural and non-natural death cases.

Determination of the associations between alcohol influence and sudden natural death represents challenges for medicolegal investigations. The aim of this study was to investigate the prevalence of alcohol influence in medicolegal autopsies. In our study of natural and non-natural deaths cases (5496 total: 4045 males, 1451 females) were examined. Blood alcohol concentrations (BACs) were detected by headspace gas chromatographic method. We investigated the alcohol-related mortality using hierarchical log-linear statistical models. Severe BACs were detected among suicidal victims in the oldest age group (>65 years) (InF = 0.442) and among the homicide victims between the age of 40-65 years (InF = 0.234). Correlations we found between manner-of-death and sex suggested that the rate of males in accidents (lnF = 0.140) and the rate of females in homicides (lnF = 0.193) were higher. It was concluded that the accurate statistical mortality database may provide a huge support for the determination of alcohol effects on human health and mortality.

The acute effect of low-dose alcohol on working memory during mental arithmetic: I. Behavioral measures and EEG theta band spectral characteristics.

OBJECTIVE AND HYPOTHESIS: Task-related EEG changes were studied during the performance of a mental arithmetic task, as influenced by low alcohol dosages with the presumption that even "social" drinking may have detrimental effect. METHODS: A mental arithmetic task was used in which addition and working memory effort was required. EEG spectra with an emphasis on the theta band, error rate and reaction time were analyzed in 5 (control, task, placebo-task, low-dose task [0.2 g/kg alcohol], high-dose task [0.4 g/kg alcohol]) conditions. Blood alcohol concentrations were measured. RESULTS: Reaction time was shortest in the placebo condition. No significant alcohol effect was seen for error rate. Task-related significant theta power increase was observed especially in the frontal area and in the left hemisphere which was reversed, although not in a significant way, by alcohol. CONCLUSION: No detrimental alcohol effect was seen on behavioral indices of task performance. However, the ethanol-induced moderate reduction of the task-related frontally dominant theta increase, probably corresponding to working memory demand, is a modest but clear electrophysiological sign of alcohol effect in this low-dose range.

The acute effect of low-dose alcohol on working memory during mental arithmetic: II. Changes of nonlinear and linear EEG-complexity in the theta band, heart rate and electrodermal activity.

OBJECTIVE AND HYPOTHESIS: Nonlinear and linear methods of EEG-complexity analysis and autonomic measures were used to characterize processes accompanying performance in a mental arithmetic task challenged by low ("social") alcohol doses. It was expected that alcohol in such doses will dampen changes of task-related EEG-synchronization in the theta band, and those of heart rate and electrodermal activity (EDA). METHODS: In the mental arithmetic task addition and working memory, effort was required. The EEG, ECG and EDA were recorded in 5 conditions: task, placebo-task, low dose-task (0.2 g/kg alcohol), high dose-task (0.4 g/kg alcohol). Omega-complexity and synchronization likelihood (SL) were computed of the theta band of the EEG. RESULTS: Task-related decrease of the Omega-complexity and increase of the SL was found in the theta frequency band. Following alcohol consumption, these changes did not develop as seen especially for SL in the anterior area, although the significant effects were elicited by task performance. Conspicuous task-evoked increases were observed for ECG and EDA which were even more enhanced by alcohol. CONCLUSIONS: Task-induced significant changes of the Omega-complexity and that of SL indicate increased synchrony in the theta band, probably corresponding to working memory effort. Both of these measures proved to be sensitive for the effect of low alcohol dose although these alcohol-elicited changes were not statistically significant. Task-induced heart rate and EDA increases were further intensified by alcohol probably indicating its activating effect on these autonomic measures in the dose range studied.